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目的探讨高浓度谷氨酸对人脑胶质瘤原代培养细胞的生长抑制和凋亡诱导作用。方法在体外对新鲜的人脑胶质瘤标本进行原代培养;不同浓度的谷氨酸作用不同时间后,进行形态学观察噻唑蓝(MTT)法检测生长抑制率;流式细胞仪Annexin V-FITC／PI双染法检测凋亡率。结果原代人脑胶质瘤细胞短期培养成功。高浓度谷氨酸对培养细胞有明显的抑制作用,并且存在一定的剂量依赖关系。当谷氨酸的浓度为25～50mmol／L时,可明显诱导培养细胞凋亡,并且凋亡率随药物浓度增加和时间延长而增高;当谷氨酸浓度达100 mmol／L以上时,细胞则主要以坏死为主。结论以酶消化法可成功对人脑胶质瘤细胞进行体外原代培养;在一定的浓度范围内,高浓度谷氨酸能明显的抑制原代培养人脑胶质瘤细胞生长,并诱导其凋亡。
Objective To investigate the effects of glutamate on growth inhibition and apoptosis induction of primary cultured human glioma cells. Methods Fresh human glioma specimens were cultured in vitro. After different concentrations of glutamic acid for different time, the growth inhibition rates were determined by MTT assay. Flow cytometry Annexin V- Apoptosis rate was detected by FITC / PI double staining. Results Primary human glioma cells were successfully cultured in short term. High concentrations of glutamate on cultured cells significantly inhibited, and there is a certain dose-dependent relationship. When glutamate concentration was 25 ~ 50mmol / L, the apoptosis of cultured cells was obviously induced, and the apoptosis rate increased with the increase of drug concentration and time. When the concentration of glutamate reached above 100 mmol / L, Mainly to necrosis. Conclusions Enzyme digestion can be used to successfully culture human glioma cells in vitro. In a certain concentration range, glutamate at high concentration can significantly inhibit the growth of primary human glioma cells and induce their growth Apoptosis.